Compounds and methods for the treatment of cardiovascular, inflammatory and immune disorders

ABSTRACT

Tetrahydrofurans, tetrahydrothiophenes, pyrrolidines and cyclopentanes are disclosed that reduce the chemotaxis and respiratory burst leading to the formation of damaging oxygen radicals of polymorphonuclear leukocytes during an inflammatory or immune response. It has been determined that 5-lipoxygenase activity, oral availability, and stability in vivo (for example, glucuronidation rate) can vary significantly among the optical isomers of the disclosed compounds.

This application is a continuation of application Ser. No. 09/023,359,filed Feb. 13, 1998 now U.S. Pat. No. 6,025,384, which is a continuationof application Ser. No. 08/454,600 filed May 31, 1995, now U.S. Pat. No.5,750,565.

FIELD OF THE INVENTION

This invention is in the area of 2,5-disubstituted tetrahydrothiophenes,tetrahydrofurans, pyrrolidines and 1,3-disubstituted cyclopentanes. Thecompounds exhibit biological activity by inhibiting the enzyme5-lipoxygenase.

BACKGROUND OF THE INVENTION

Leukotrienes are potent local mediators, playing a major role ininflammatory and allergic responses, including arthritis, asthma,psoriasis, and thrombotic disease. Leukotrienes are straight chaineicosanoids produced by the oxidation of arachidonic acid bylipoxygenases. Arachidonic acid is oxidized by 5-lipoxygenase to thehydroperoxide 5-hydroperoxy-eicosatetraenoic acid (5-HPETE), that isconverted to leukotriene A₄, that in turn can be converted toleukotriene B₄, C₄, or D₄. The slow-reacting substance of anaphylaxis isnow known to be a mixture of leukotrienes C₄, D₄, and E₄, all of whichare potent bronchoconstrictors. There has been a research effort todevelop specific receptor antagonists or inhibitors of leukotrienebiosynthesis, to prevent or minimize pathogenic inflammatory responsesmediated by these compounds.

European Patent Application Nos. 90117171.0 and 901170171.0 discloseindole, benzofuran, and benzothiophene lipoxygenase inhibitingcompounds.

Recently, it was reported that the tetrahydrothiophene derivative ofL-652,731, trans-2,5-bis-(3,4,5-trimethoxyphenyl)tetrahydrothiophene(L-653,150), is a potent PAF antagonist and a moderate inhibitor of5-lipoxygenase. It has been disclosed that certain 2,5-diaryltetrahydrothiophenes are PAF antagonists and leukotriene synthesisinhibitors. (Biftu, et al., Abstr. of 6^(th) Int. Conf. onProstaplandins and Related Compounds, Jun. 3-6, 1986, Florence, Italy;U.S. Pat. No. 4,757,084 to Biftu); WO 92/15294; WO 94/01430; WO94/04537; and WO 94/06790.

WO 92/13848 discloses a class of racemic lipoxygenase inhibitinghydroxamic acid and N-hydroxyurea derivatives of the structure

wherein R¹ is hydrogen, alkyl, alkenyl, amino or substituted amino, R⁴is hydrogen, a pharmaceutically acceptable cation, aroyl or alkoyl, A isalkylene or alkenylene, X is oxygen or sulfur, each Y is hydrogen, halo,cyano, alkyl, alkoxy, alkylthio, alkenyl, alkoxyalkyl, cycloalkyl,cycloalkyl, aryl, aryloxy, arylalkyl, arylalkenyl, arylalkoxy orsubstituted aryl, Z is oxygen or sulfur, m is 0 or 1, n is 1 to 5 and pis 2 to 6, inhibit the enzyme lipoxygenase.

Given the significant number of pathological immune and inflammatoryresponses that are mediated by 5-lipoxygenase, there remains a need toidentify new compounds and compositions that inhibit this enzyme.

Therefore, it is an object of the present invention to provide compoundsthat reduce the chemotaxis and respiratory burst leading to theformation of damaging oxygen radicals during an inflammatory or immuneresponse.

It is another object of the present invention to provide pharmaceuticalcompositions for the treatment of pathological immune or inflammatorydisorders mediated by 5-lipoxygenase.

It is another object of the present invention to provide a method forthe treatment of pathological immune or inflammatory disorders mediatedby products of 5-lipoxygenase.

SUMMARY OF THE INVENTION

Compounds of Formula I are provided

wherein:

Ar is an aryl or heteroaryl group that is optionally substituted,preferably with halo (including but not limited to fluoro), lower alkoxy(including methoxy), lower aryloxy (including phenoxy), W, cyano, or R³;

m is 0 or 1;

n is 1-6;

W is independently —AN(OM)C(O)N(R³)R⁴, —N(OM)C(O)N(R³)R⁴,—AN(R³)C(O)N(OM)R⁴, —N(R³)C(O)N(OM)R⁴, —AN(OM)C(O)R⁴, —N(OM)C(O)R⁴—,AC(O)N(OM)R⁴, —C(O)N(OM)R⁴, or —C(O)NHA; A in lower alkyl, loweralkenyl, lower alkynyl, alkylaryl or arylalkyl groups, wherein one ormore carbons optionally can be replaced by O, N, or S (with valencecompleted with hydrogen or oxygen as necessary), however, —Y—A—, —A—, or—AW— should not include two adjacent heteroatoms (i.e. —O—O—, —S—S—,—O—S—, etc.) (in one embodiment, lower alkyl is a branched alkyl groupsuch as —(CH₂)₀C(alkyl)H—, wherein n is 1-5, and specifically—(CH)₂C(CH₃)H—, or lower alkynyl of the formula C≡C—CH(alkyl)-,including —C≡C—CH(CH₃)—);

M is hydrogen, a pharmaceutically acceptable cation, or a metabolicallycleavable leaving group;

X is O, S, S(O), S(O)₂, NR³, or CHR⁵;

Y is O, S, S(O), S(O)₂, NR₃, or CHR⁵;

Z is O, SO S(O), S(O)₂, NR³;

R¹ and R² are independently hydrogen, lower alkyl including methyl,cyclopropylmethyl, ethyl, isopropyl, butyl, pentyl hexyl, and C₃₋₈cycloalkyl, for example, cyclopentyl; halo lower alkyl, for example,trifluoromethyl; halo, for example fluoro; and —COOH;

R³ and R⁴ are independently hydrogen or alkyl, alkenyl, alkynyl, aryl,arylalkyl, alkylaryl, C₁₋₆ alkoxy-C₁₋₁₀ alkyl, C₁₋₆ alkylthio-C₁₋₁₀alkyl, heteroaryl, or heteroarylalkyl-;

R⁵ is hydrogen, lower alkyl, lower alkenyl, lower alkynyl, arylalkyl,alkyaryl, —AN(OM)C(O)N(R³)R⁴, —AN(R³)C(O)N(OM)R⁴, —AN(OM)C(O)R⁴,—AC(O)N(OM)R⁴, —AS(O)_(x)R³, —AS(O)_(a)CH₂C(O)R³, —AS(O)_(a)CH₂CR(OH)R⁴,or —AC(O)NHR³, wherein x is 0-2;

The Ar group, in one embodiment, is selected from the group consistingof phenyl, trimethoxyphenyl, dimethoxyphenyl, fluorophenyl (specifically4-fluorophenyl), difluorophenyl, pyridyl, dimethoxypyridyl, quinolinyl,furyl, imidarolyl, and thienyl groups.

Nonlimiting examples of preferred compounds are:

wherein R¹⁰ is halogen, —CN, hydrogen, lower alkyl, lower alkenyl, loweralkynyl, lower alkoxy, or lower aryloxy.

These compounds in general reduce the chemotaxis and respiratory burstleading to the formation of damaging oxygen radicals ofpolymorphonuclear leukocytes during an inflammatory or immune response.The compounds exhibit this biological activity by inhibiting the enzyme5-lipoxygenase.

Another embodiment of the present invention is a pharmaceuticalcomposition that includes an effective amount of a compound of Formula Ior its pharmaceutically acceptable salt or derivative in combinationwith a pharmaceutically acceptable carrier for any of the disordersdescribed herein.

A method to treat disorders mediated by 5-lipoxygenase is alsodisclosed, that includes administering an effective amount of one ormore of the above-identified compounds or a pharmaceutically acceptablesalt or derivative thereof, optionally in a pharmaceutically acceptablecarrier.

It has been surprisingly determined that 5-lipoxygenase activity, oralavailability, and stability in vivo (for example, glucuronidation rate)can vary significantly among the optical isomers of the disclosedcompounds. Therefore, in one embodiment of the invention, the compoundis administered in an enantiomerically enriched form, i.e.,substantially in the form of one isomer.

Examples of immune, allergic and cardiovascular disorders includegeneral inflammation, cardiovascular disorders including hypertension,skeletal-muscular disorders, osteoarthritis, gout, asthma, lung edema,adult respiratory distress syndrome, pain, aggregation of platelets,shock, rheumatoid arthritis, juvenile rheumatoid arthritis, psoriaticarthritis, psoriasis, autoimmune uveitis, allergic encephalomyelitis,systemic lupus erythematosis, acute necrotizing hemorrhagicencephalopathy, idiopathic thrombocytopenia, polychondritis, chronicactive hepatitis, idiopathic sprue, Crohn's disease, Gravesophthalmopathy, primary biliary cirrhosis, uveitis posterior,interstitial lung fibrosis; allergic asthma; and inappropriate allergicresponses to environmental stimuli such as poison ivy, pollen, insectstings and certain foods, including atopic dermatitis and contactdermatitis.

The compounds disclosed herein can also be used as research tools tostudy biological pathways involving leukotrienes.

The following are nonlimiting examples of compounds that fall withinFormula I. These examples are merely exemplary, and are not intended tolimit the scope of the invention.

trans-2-(3,4,5-trimethoxyphenoxymethyl)-5-[4-N′-methyl-N′-hydroxyureidyl)butyl]tetrahydrofuran

trans-2-(3,4,5-trimethoxyphenoxymethyl)-5-[4-N′-methyl-N′-hydroxyureidyl)but-1-ynyl]tetrahydrofuran

trans-2-(4-fluorophenoxymethyl)-5-[4-N′-methyl-N′-hydroxyureidyl)butyl]tetrahydrofuran

trans-2-(4-fluorophenoxymethyl)-5-[4-N′-methyl-N′-hydroxyureidyl)but-1-ynyl]tetrahydrofuran

trans-2-(4-fluorophenoxymethyl)-5-[4-N′-butyl-N′-hydroxyureidyl)butyl]tetrahydrofuran

trans-2-(4-fluorophenoxymethyl)-5-[4-N′-butyl-N′-hydroxyureidyl)but-1-ynyl]tetrahydrofuran

trans-2-(3,4,5-trimethoxyphenoxymethyl)-5-[4-N′methyl-N-hydroxyureidyl)butyl]tetrahydrofuran

trans-2-(3,4,5-trimethoxyphenoxymethyl)-5-[4-N′-methyl-N-hydroxyureidyl)but-1-ynyl]tetrahydrofuran

trans-2-(4-fluorophenoxymethyl)-5-[4-N′-methyl-N-hydroxyureidyl)butyl]tetrahydrofuran

trans-2-(4-flourophenoxymethyl)-5-[4-N′-methyl-N-hydroxyureidyl)but-1-ynyl]tetrahydrofuran

trans-2-(4-fluorophenoxymethyl)-5-[4-N′-butyl-N-hydroxyureidyl)butyl]tetrahydrofuran

trans-2-(4-fluorophenoxymethyl)-5-[4-N′-butyl-N-hydroxyureidyl)but-1-ynyl]tetrahydrofuran

trans-2-(3,4,5-trimethoxyphenoxymethyl)-5-[4-N′-methyl-N′-hydroxyureidyl)butyl]tetrahydrothiophene

trans-2-(3,4,5-trimethoxyphenoxymethyl)-5-[4-N′-methyl-N′-hydroxyureidyl)but-1-ynyl]tetrahydrothiophene

trans-2-(4-fluorophenoxymethyl)-5-[4-N′-methyl-N′-hydroxyureidyl)butyl]tetrahydrothiophene

trans-2-(4-fluorophenoxymethyl)-5-[4-N′-methyl-N′-hydroxyureidyl)but-1-ynyl]tetrahydrothiophene

trans-2-(4-fluorophenoxymethyl)-5-[4-N′-butyl-N′-hydroxyureidyl)butyl]tetrahydrothiophene

trans-2-(4-fluorophenoxymethyl)-5-[4-N′-butyl-N′-hydroxyureidyl)but-1-ynyl]tetrahydrothiophene

trans-2-(3,4,5-trimethoxyphenoxymethyl)-5-[4-N′-methyl-N-hydroxyureidyl)butyl]tetrahydrothiophene

trans-2-(3,4,5-trimethoxyphenoxymethyl)-5-[4-N′-methyl-N-hydroxyureidyl)but-1-ynyl]tetrahydrothiophene

trans-2-(4-fluorophenoxymethyl)-5-[4-N′-methyl-N-hydroxyureidyl)butyl]tetrahydrothiophene

trans-2-(4-fluorophenoxymethyl)-5-[4-N′-methyl-N-hydroxyreidyl)but-1-ynyl]tetrahydrofuran

trans-2-(4-flourophenoxymethyl)-5-[4-N′-hydroxyureidyl)butyl]tetrahydrothiophene

trans-2-(4-fluorophenoxymethyl)-5-[4-N′-butyl-N-hydroxyureidyl)but-1-ynyl]tetrahydrothiophene

Further nonlimiting examples of other compounds that fall within FormulaI are set forth below in Table 1.

TABLE 1 —(Y)_(m)—W —(Y)_(m)—W

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is an illustration of one process for the synthesis of2S,5S-trans-2-(4-fluorophenoxymethyl)-5-(4-N-hydroxynreidyl-1-butynyl)tetrahydrofuran(compound 1) and2S,5R-trans-2-(4-fluorophenoxymethyl)-5-(4-N-hydroxyureidylbutyl)tetrahydrofuran (compound 2).

FIG. 2 is a graph of the rate of glucuronidation of compounds 1,3,4, and6 (as illustrated in Table 2) as measured in percent metabolite versustime (hours).

DETAILED DESCRIPTION OF THE INVENTION I. Description and Synthesis ofthe Compounds

A. Compounds

As used herein, the term “enantiomerically enriched” refers to acompound in the form of at least approximately 95%, and preferablyapproximately 97%, 98%, 99%, or 100% of a single enantiomer of thatcompound.

The term alkyl, as used herein, unless otherwise specified, refers to asaturated straight, branched, or cyclic. hydrocarbon of C₁ to C₁₀ andspecifically includes methyl, ethyl, propyl, isopropyl, butyl, isobutyl,t-butyl, pentyl, cyclopentyl, isopentyl, neopentyl, hexyl, isohexyl,cyclohexyl, 3-methylpentyl, 2,2-dimethylbutyl, and 2,3-dimethylbutyl.The alkyl group can be optionally substituted with any appropriategroup, including but not limited to R³ or one or more moleties selectedfrom the group consisting of halo, hydroxyl, amino, alkylamino,arylamino, alkoxy, aryloxy, nitro, cyano, sulfonic acid, sulfate,phosphonic acid, phosphate, or phosphonate, either unprotected, orprotected as necessary, as known to those skilled in the art, forexample, as taught in Greene, et al., “Protective Groups in OrganicSynthesis,” John Wiley and Sons, Second Edition, 1991.

The term halo, as used herein, refers to chloro, fluoro, iodo, or bromo.

The term lower alkyl, as used herein, and unless otherwise specified,refers to a C₁ to C₆ saturated straight, branched, or cyclic (in thecase of C₅₋₆) hydrocarbon, and specifically includes methyl, ethyl,propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, cyclopentyl,isopentyl, neopentyl, hexyl, isohexyl, cyclohexyl, 3-methylpentyl,2,2-dimethylbutyl, and 2,3-dimethylbutyl, optionally substituted asdescribed above for the alkyl groups.

The term alkenyl, as referred to herein, and unless otherwise specified,refers to a straight, branched, or cyclic (in the case of C₅₋₆)hydrocarbon of C₂ to C₁₀ with at least one double bond,optionallysubstituted as described above.

The term lower alkenyl, as referred to herein, and unless otherwisespecified, refers to an alkenyl group of C₂ to C₆, and specificallyincludes vinyl and allyl.

The term lower alkylamino refers to an amino group that has one or twolower alkyl substituents.

The term alkynyl, as referred to herein, and unless otherwise specified,refers to a C₂ to C₁₀ straight or branched hydrocarbon with at least onetriple bond, optionally substituted as described above. The term loweralkynyl, as referred to herein, and unless otherwise specified, refersto a C₂ to C₆ alkynyl group, specifically including acetylenyl,propynyl, and —C≡C—CH(alkyl)-, including —C≡C—CH(CH₃)—.

The term aryl, as used herein, and unless otherwise specified, refers tophenyl, biphenyl, or napthyl, and preferably phenyl. The aryl group canbe optionally substituted with any suitable group, including but notlimited to one or more moieties selected from the group consisting ofhalo, hydroxyl, amino, alkylamino, arylamino, alkoxy, aryloxy, nitro,cyano, sulfonic acid, sulfate, phosphoric acid, phosphate, orphosphonate, either unprotected, or protected as necessary, as known tothose skilled in the art, for example, as taught in Greene, et al.,“Protective Groups in Organic Synthesis,” John Wiley and Sons, SecondEdition, 1991, and preferably with halo (including but not limited tofluoro), lower alkoxy (including methoxy), lower aryloxy (includingphenoxy), W, cyano, or R³.

The term haloalkyl, haloalkenyl, or haloalkynyl refers to alkyl,alkenyl, or alkynyl group in which at least one of the hydrogens in thegroup has been replaced with a halogen atom.

The term heteroaryl, heterocycle or heteroaromatic, as used herein,refers to an aromatic moiety that includes at least one sulfur, oxygen,or nitrogen in the aromatic ring, which can optionally be substituted asdescribed above for the aryl groups. Non-limiting examples are pyrryl,furyl, pyridyl, 1,2,4-thiadiazolyl, pyrimidyl, thienyl, isothiazolyl,imidazolyl, tetrazolyl, pyrazinyl, pyrimidyl, quinolyl, isoquinolyl,benzothienyl, isobenzofuryl, pyrazolyl, indolyl, purinyl, carbazolyl,benzimidazolyl, and isoxazolyl.

The term arylalkyl refers to an aryl group with an alkyl substituent.

The term alkylaryl refers to an alkyl group that has an arylsubstituent.

The term organic or inorganic anion refers to an organic or inorganicmoiety that carries a negative charge and can be used as the negativeportion of a salt.

The term “pharmaceutically acceptable cation” refers to an organic orinorganic moiety that carries a positive charge and that can beadministered in association with a pharmaceutical agent, for example, asa countercation in a salt.

Pharmaceutically acceptable cations are known to those of skill in theart, and include but are not limited to sodium, potassium, andquaternary amine.

The term metabolically cleavable leaving group% refers to a moiety thatcan be cleaved in vivo from the molecule to which it is attached, andincludes but is not limited to an organic or inorganic anion, apharmaceutically acceptable cation, acyl (for example (alkyl)C(O),including acetyl, propionyl, and butyryl), alkyl, phosphate, sulfate andsulfonate.

The term 5-lipoxygenase inhibitor refers to a compound that inhibits theenzyme at 30 μM or lower in a broken cell system.

The term pharmaceutically active derivative refers to any compound thatupon administration to the recipient, is capable of providing directlyor indirectly, the compounds disclosed herein.

The 2,5-disubstituted tetrahydrothiophenes, tetrahydrofurans andpyrrolidines, as well as the 1,3-disubstituted cyclopentanes describedherein inhibit the enzyme 5-lipoxygenase and are thus useful in thetreatment of humans who have immune allergic or cardiovascular disordersthat are mediated by 5-lipoxygenase.

B. Stereochemistry

It has been surprisingly determined that 5-lipoxygenase activity, oralavailability, and stability in vivo (for example, glucuronidation rate)can vary significantly among the optical isomers of the disclosedcompounds. Therefore, in a preferred embodiment, the active compound orits precursor is administered in an enantiomerically enriched form,i.e., substantially in the form of one isomer. The preferred enantiomeris easily determined by evaluating the various possible enantiomers inselected biological assays, for example, those described in detailherein.

The 2,5-disubstituted tetrahydrofurans, etrahydrothiophenes, andpyrrolidines exhibit a number of stereochemical configurations. Carbonatoms 2 and 5 in the center ring are chiral, and thus the center ringexists at a minimum as a diastereomeric pair. Each diastereomer existsas a set of enantiomers. Therefore, based on the chiral C₂ and C₅ atomsalone, the compound is a mixture of four enantiomers.

If non-hydrogen substituents are located on carbon atoms 3 and 4 in thecenter ring, then the C₃ and C₄ atoms are also chiral, and can alsoexist as a diastereomeric pair, that is also a mixture of fourenantiomers.

The 1,3-cyclopentanes disclosed herein also exhibit a number ofstereochemical configurations. Carbon atoms 1 and 3 in the center ringare chiral, and thus the center ring exists at a minimum as adiastereomeric pair. Each diastereomer exists as a set of enantiomers.Therefore, based on the chiral C₁ and C₃ atoms alone, the compound is amixture of four enantiomers.

If non-hydrogen substituents are located on carbon atoms 4 and 5 in thecenter ring, then the C₄ and C₅ atoms are also chiral, and can alsoexist as a diastereomeric pair, that is also a mixture of fourenantiomers.

One of ordinary skill in the art can easily synthesize and separate theenantiomers of the disclosed compounds using chiral reagents and knownprocedures, and can evaluate the biological activity of the isolatedenantiomer using methods disclosed herein or otherwise known. Throughthe use of chiral NMR shift reagents, polarimetry, or chiral HPLC, theoptical enrichment of the compound can be determined.

Classical methods of resolution include a variety of physical andchemical techniques. For example, if the compound is basic, one can usechiral acids that form diastereomeric derivatives that may possesssignificantly different solubility properties. Non-limiting examples ofchiral acids include malic acid, mandelic acid, dibenzoyl tartaric acid,3-bromocamphor-8-sulfonic acid, 10-camphorsulfonic acid, anddi-p-toluoyltartaric acid. Similarly, acylation of a free hydroxyl groupwith a chiral acid also results in the formation of diastereomericderivatives whose physical properties may differ sufficiently to permitseparation.

Enantiomerically pure or enriched compounds can also be obtained bypassing the racemic mixture through a chromatographic column that hasbeen designed for chiral separations, or by enzymatic resolution ofappropriately modified substrates.

C. Syntheses of Active Compounds

The 2,5-disubstituted tetrahydrofurans, tetrahydrothiophenes, andpyrrolidines disclosed herein can be prepared in a variety of ways knownto those skilled in the art, including by methods disclosed by, orroutine modifications of the methods disclosed by, Whittaker et al,Synlett, 1993 pp 111, Biorg. Med. Lett., 1993 pp 1499; Achiwa et al.,Chem. Pharm. Bull., 1989, pp. 1969. These compounds can prepared in bothracemic and enantiomerically enriched forms.

1,3-Disubstituted cyclopentanes can be prepared using the procedure ofGraham, et al. (1,3-Diaryl Cyclopentanes: A New Class of Potent PAFReceptor Antagonists. 197^(th) ACS National Meeting, Dallas, Tex., Apr.9-14, 1989, Division of Medicinal Chemistry, poster no. 25 (abstract)),or by other known methods.

A general procedure for preparing a hydroxyurea is shown below in Scheme1.

General procedures for preparing reverse hydroxyureas are shown inScheme. 2

A general procedure for preparing a hydroxamic acid is shown in Scheme3.

A general procedure for preparing a reverse hydroxamic acid is shown inScheme 4.

The following working examples, depicted in FIG. 1, are merelyillustrative, and are not intended to limit the scope of the invention.Other compounds falling within the disclosed formula can be prepared byone of skill in the art by modification of the scheme below or by otherknown methods.

EXAMPLE 1 Preparation of2S,5S-trans-2-(4-Fluorophenoxymethyl)-5-(4-N-hydroxyureidyl-1-butynyl)tetrahydrofuran(Compound 1, FIG. 1) and2S,5R-trans-2-(4-Fluorophenoxymethyl)-5-(4-N-hydroxyureidylbutyl)tetrahydrofuran(Compound 2, FIG. 1) Preparation of4S-(4-Fluorophenoxymethyl)-gammabutyrolactone (Compound 101, FIG. 1)

To a stirred THF (10 mL) solution of (S)-gamma-butyrolactone (1.0 g,8.61 mmol), 4-fluorophenol (1.16, g, 10.35 mmol), and triphenylphosphine(2.49 g, 9.49 mmol) was added diisopropoxyl azodicarboxylate (1.87 μL,9.46 mmol) dropwise. Alter addition, the reaction mixture was stirred at80° C. for 16 hours. The solvent was removed and the product wasseparated by flash column chromatography (silica, 2:1 hexane/ethylacetate) (1.38 g, 76.3%). ¹H NMR (CDCl₃); 2.27 (m, 1H); 2.42 (m, 1H);2.60 (m, 1H); 4.04 (m, 1H); 4.15 (m, 1H); 4.85 (m, 1H); 6.84 (m, 2H);6.98 (m, 2H).

Preparation of 2S-(4-Fluorophenoxymethyl)-5-hydroxy-tetrahydrofuran(Compound 102, FIG. 1).

To a stirred solution of lactone 101 (1.38 g, 27.22 mmol) in dry toluene(24 mL) at −78° C. was added a 1.5 M toluene solution of DIBAL (6.76 mL,10.13 mmol) dropwise. The reaction mixture was stirred at −78° C. for 2hours. The reaction was quenched through the addition of methanol (1.7ml) while maintaining a temperature of <−60° C. The mixture was warmedto −20° C. followed by the addition of saturated aqueous potassiumsodium tartrate solution (10 mL) while maintaining the reactiontemperature between −10 and 0° C. The reaction mixture was stirred atroom temperature overnight and then the two phases were separated. Theaqueous layer was extracted with ethyl acetate. The combined organiclayers were washed with water, saturated NaCl solution, and thenconcentrated in vacuo to leave an oil which was purified by flash columnchromatography (silica, 1:1 hexane/ethyl acetate) (1.41 g, 101%). ¹H NM(CDCl₃); 1.80 (m, 1H); 2.05 (m, 2H); 2.26 (m, 1H); 3.9.3 (m, 2H); 4.04(m, 2H); 4.47 (m, 0.5H); 4.61 (m, 0.5H); 5.57 (m, 0.5H); 5.66 (m, 0.5H);6.88 (m, 2H); 6.98 (m, 2H).

Preparation of2S-(4-Fluorophenoxymethyl)-5-(t-butyldimethysiloxy)tetrahydrofuran(Compounds 103, FIG. 1)

To a stirred solution of lactol 102 (1.41 g, 6.65 mmol) in methylenechloride (25 mL) was added imidazole (498.1 mg, 7.32 mmol) and TBDMSchloride (1.10 g, 7.32 mmol). The reaction mixture was stirred at roomtemperature overnight and then the reaction was filtered and thefiltrate was concentrated. The crude product was purified by flashcolumn chromatography using 9:1 hexane/ethyl acetate as a solvent togive a colorless oil which is a mixture of two diasteriomers (ca. 2:1)(1.22 g, 56.2%). ¹H NMR (CDCl₃); 0.11 (s, 6H); 0.90 (s, 9H); 1.80-2.10(m, 3H); 2.22 (m, 1H); 3.91 (m, 2H); 4.38 (m, 0.33H); 4.50 (m, 0.67H);5.52 (m, 0.33H); 5.59 (m, 0.67H); 6.86 (m, 2H); 6.96 (m, 2H);

Preparation of2S,5S-trans-2-(4-Flourophenoxymethyl)-5-(4-t-butyldimethysiloxy-1-butynyl)tetrahydrofuran(Compounds 104, FIG. 1) and2S,5R-cis-2-(4-Fluorophenoxymethyl)-5-(4-t-butyldimethysiloxy-1-butynyl)tetrahydrofuran(Compounds 105, FIG. 1)

To a stirred solution of 103 (720 mg, 2.21 mmol) in dry methylenechloride (10 mL), cooled to −78° C. was added TMS bromide (349.8 μL,2.65 mmol). The reaction mixture was stirred at −78° C. for 4 hours. Ina separate flask containing 4-t-butyldimethylsiloxy-1-butyne (812.8 mg,4.42 mmol) and THF (10 mL) was added n-butyllithium (2.5M in hexane,2.65 mL, 6.63 mmol). After 30 minutes, this was transferred by cannulato the solution from above. After two hours, the reaction was quenchedthrough the addition of saturated aq. ammonium chloride solution andextracted with methylene chloride, dried over MgSO4, filtered andconcentrated. Flash column chromatography (silica, 95:5 hexane/ethylacetate) yielded two products, trans compound 104 (210 mg) and ciscompound 105 (160 mg), and the mixture of these two compounds (50 mg).The total yield is 48.5%. ¹H NMR (CDCl₃); 104: 0.10 (s, 6H); 0.91 (s,9H); 1.87 (m, 1H); 2.01 (m, 1H); 2.22 (m, 2H); 2.43 (t, 2H); 3.72 (t,2H); 3.92 (d, 2H); 4.47 (m, 1H); 4.73 (m, 1H); 6.86 (m, 2H); 6.95 (t,2H). 105: 0.09 (s, 6H); 0.90(s, 9H); 1.92-2.20 (m, 4H); 2.42 (m, 2H);3.70 (t, 2H); 3.92 (m, 1H); 4.07 (m, 1H); 4.29 (m, 13); 4.62 (m, 1H);6.86 (m, 2H); 6.96 (t, 2H.

In the preparation of compounds 104 and 105, oxygen protecting groupsknown to those skilled in the art other than 4-t-butyldimethysilyl canbe used as desired.

In order to determine the stereochemistry of this molecule, a NOEdifference experiment was carried out for both 104 and 105. In the NOEdifference experiment of 104, the multiplet at 4.73 ppm was irradiatedwith a very low rf decoupling pulse and the data work-up was done so asto only measure the presence of an increase in signal. This wouldrepresent a positive NOE effect and would indicate the close spacialrelationship of these protons. In this experiment, a NOE was found forthe multiplet at 2.22 ppm which are furan ring protons. When themultiplet at 4.47 ppm was irradiated with a very low rf decoupling pulseand the data work-up was done so as to only measure the presence of anincreased in signal. A NOE was found for the multiplet at 2.22 ppm whichare furan ring protons. Another NOE was also seen for the protons at3.92 ppm which are the protons on the methylene next to this multiplet,indicating that this multiplet represents the proton next to themethylene.

In the NOE difference experiment of 105, the triplet at 4.62 ppm wasirradiated with a very low rf decoupling pulse and the data work-up wasdone so as to only measure the presence of an increase in signal. Thiswould represent a positive NOE effect and would indicate the closespacial relationship of these protons. In this experiment, a NOE wasfound for the multiplet at 4.29 ppm which is the other methine furanproton. Another NOE was also seen for the multiplet at 2.17 ppm whichare furan protons. When the multiplet at 4.29 ppm was irradiated with avery low rf decoupling pulse and the data work-up was done so as to onlymeasure the presence of an increase in signal. A NOE was found for thetriplet at 4.62 ppm which is the other methine furan proton. Another NOEwas seen for the protons at 3.92 and 4.07 ppm which are the protons onthe methylene next to this multiplet, indicating that this multipletrepresents the proton next to the methylene. Another NOE was also seenfor the multiplet at 2.11 ppm which are furan protons.

Preparation of2S,5S-trans-2-(4-Fluorophenoxymethyl)-5-(4-t-hydroxy-1-butyl)tetrahydrofuran(Compounds 106, FIG. 1)

To a stirred solution of 104 (210 mg, 0. 54 mmol) in, THF (1.4 mL),cooled in an ice bath, was added tetrabutyl ammonium fluoride (420.3 mg,1.61 mmol). The ice bath was removed and the reaction was stirred atroom temperature for 1 hour. The solvent was removed and the product wasseparated by flash column chromatography (silica, 1:1 hexane/ethylacetate) (124 mg, 83.2%). ¹H NMR (CDCl₃); 1.88 (m, 1H); 2.02 (m, 1H);2.25 (m, 2H); 2.50 (m, 2H); 3.72 (t, 2H); 3.93 (d, 2H); 4.48 (m, 1H);4.76 (m, 1H); 6.84 (m, 2H); 6.96 (t, 2H).

Preparation of2S,5S-trans-2-(Fluorophenoxymethyl)-5-(4-N,O-bisphenoxycarbonylhydroxylamino-1-butynyl)tertrahydrofuran(Compound 107, FIG. 1)

To a cooled (ice bath) solution compound 106 (124.0 mg, 0.45 mmol),triphenylphosphine (128.9 mg., 0.49 mmol) andN,O-bisphenoxycarbonylhydroxylamine (147.3 mg, 0.54 mmol) in THF (5 mL)was added diisopropoxylazodicarboxylate (94.1 μL, 0.48 mmol). The icebath was removed and the reaction was warmed to room temperature andstirred at room temperature for 30 minutes. The solvent was removed andthe product was purified by flash column chromatography (silica, 4:1hexane/ethyl acetate) (195 mg, 82.0%). ¹H NMR(CDCl₃); 1.85 (m, 1H); 2.03(m, 1H); 2.22 (m, 2H); 2.75 (m, 2H); 3.92 (d, 2H); 4.05 (m, 2H); 4.47(m, 1H); 4.76 (m, 1H); 6.84 (m, 2H); 6.95 (m, 2H); 7.26 (m, 5H); 7.41(m, 5H).

Preparation of2S,5S-trans-2-(4-Fluorophenoxymethyl)-5-(4-N-hydroxyureidyl-1-butynyl)tetrahydrofuran(Compound 1, FIG. 1)

In a screw top vessel was placed MR. at −78° C. (approximately 1-2 mL).Compound 107 (195.0 mg, 0.37 mmol), predissolved in 20 mL methanol, wasadded to this cold liquid nitrogen. The vessel was sealed and the dryice bath was removed. The reaction mixture was stirred at roomtemperature for 16 hours. The reaction mixture was cooled again by dryice bath and the pressure was released. The vessel was opened and thesolvent was removed. The product was isolated by flash columnchromatography using ethyl acetate as a solvent to provide a solid (108mg. 91.7%). ¹H NMR (CDCl₃); 1.84 (m, 1H); 2,01 (m, 1H); 2.2,2 (m, 2H);2.55 (t, 2H); 3.75 (t, 2H); 3.94(m, 2H); 4.48 (m, 1H); 4.74 (t, 1H);5.25 (bs, 2H); 6.86 (m, 2H); 6.98 (m, 2H).

Preparation of2S,5S-trans-2-(4-Fluorophenoxymethyl)-5-(4-N-hydroxyureidyl-1-butynyl)tetrahydrofuran(2)

Compound 1 (75 mg, 0.23 mmol) was dissolved in 2 mL of ethyl acetate andthen Pd/C (10%) (15 mg) was added and hydrogenated at balloon pressurefor 16 hours. The reaction was filtered and the filtrate wasconcentrated. The product was isolated by flash column chromatographyusing ethyl acetate as solvent (70 mg, 92.2%). ¹H NMR (CDCl₃); 1.50-1.70(m, 8H); 2.10 (m, 2H); 3.58 (m, 2H); 3.91 (m, 2H); 4.08 (m, 1H); 4.40(m, 1H); 5.15(bs, 2H); 6.87 (m, 2H); 6.97 (t, 2H); 7.40 (bs, 1H).

II. Pharmaceutical Compositions

Humans, equines, canines, bovines and other animals, and in particular,mammals, suffering from inflammatory diseases, and in particular,disorders mediated by 5-lipoxygenase, can be treated by administering tothe patient an effective amount of one or more of the above-identifiedcompounds or a pharmaceutically acceptable derivative or salt thereof ina pharmaceutically acceptable carrier or diluent. The active materialscan be administered by any appropriate route, for example, orally,parenterally, intravenously, intracermally, subcutaneously, ortopically, in liquid, cream, gel, or solid form, or by aerosol form.

As used herein, the term pharmaceutically acceptable salts or complexesrefers to salts or complexes that retain the desired biological activityof the above-identified compounds and exhibit minimal undesiredtoxicological effects. Non-limiting examples of such salts are (a) acidaddition salts formed with inorganic acids (for example, hydrochloricacid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, andthe like), and salts formed with organic acids such as acetic acid,oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic acid,benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic acid,naphthalenesulfonic acid, naphthalenedisulfonic acid, andpolygalacturonic acid; (b) base addition salts formed with metal cationssuch as zinc, calcium, bismuth, barium, magnesium, aluminum, copper,cobalt, nickel, cadmium, sodium, potassium, and the like, or with acation formed from ammonia, N,N-dibenzylethylene-diamine, D-glucosamine,tetraothylammonium, or ethylenediamine; or (c) combinations of (a) and(b); e.g., a zinc tannate salt or the like. The compounds can also beadministered as pharmaceutically acceptable quaternary salts known bythose skilled in the art, which specifically include the quaternaryammonium salt of the formula —NR⁺Z⁻, wherein R is alkyl or benzyl, and Zis a counterion, including chloride, bromide, iodide, —O-alkyl,toluenesulfonate, methylsulfonate, sulfonate, phosphate, or carboxylate(such as benzoate, succinate, acetate, glycolate, maleate, malate,malate, citrate, tartrate, ascorbate, benzoate, cinnamoate, mandeloate,benzyloate, and diphenylacetate.

The active compound is included in the pharmaceutically acceptablecarrier or diluent in an amount sufficient to deliver to a patient atherapeutically effective amount without causing serious toxic effectsin the patient treated. A preferred dose of the active compound for allof the above-mentioned conditions is in the range from about 10 ng/kg to300 mg/kg, preferably 0.1 to 100 mg/kg per day, more generally 0.5 toabout 25 mg per kilogram body weight of the recipient per day. Apreferred dosage for cardiovascular indications is in the range 10 ng/kgto 20 mg/kg. A typical topical dosage will range from 0.01-3% wt/wt in asuitable carrier. The effective dosage range of the pharmaceuticallyacceptable derivatives can be calculated based on the weight of theparent compound to be delivered. If the derivative exhibits activity initself, the effective dosage can be estimated as above using the weightof the derivative, or by other means known to those skilled in the art.

The compound is conveniently administered in any suitable unit dosageform, including but not limited to one containing 1 to 3000 mg,preferably 5 to 500 mg of active ingredient per unit dosage form. A oraldosage of 25-250 mg is usually convenient.

The active ingredient is preferably administrated to achieve peak plasmaconcentrations of the active compound of about 0.00001-30 mM, preferablyabout 0.1-30 μM.

The concentration of active compound in the drug composition will dependon absorption, distribution, inactivation, and excretion rates of thedrug as well as other factors known to those of skill in the art. It isto be noted that dosage values will also vary with the severity of thecondition to be alleviated. It is to be further understood that for anyparticular subject, specific dosage regimens should be adjusted overtime according to the individual need and the professional judgment ofthe person administering or supervising the administration of thecompositions, and that the concentration ranges set forth herein areexemplary only and are not intended to limit the scope or practice ofthe claimed composition. The active ingredient may be administered atonce, or may be divided into a number of smaller doses to beadministered at varying intervals of time.

Oral compositions will generally include an inert diluent or an ediblecarrier. They may be enclosed in gelatin capsules or compressed intotablets. For the purpose of oral therapeutic administration, the activecompound can be incorporated with excipients and used in the form oftablets, troches, or capsules.

Pharmaceutically compatible binding agents, and/or adjuvant materialscan be included as part of the composition.

The tablets, mills, capsules, troches and the like can contain any ofthe following ingredients, or compounds of a similar nature: a bindersuch as microcrystalline cellulose, gum tragacanth or gelatin; anexcipient such as starch or lactose, a dispersing agent such as alginicacid, Primogel, or corn starch; a lubricant such as magnesium stearateor Sterotes; a glidant such as colloidal silicon dioxide; a sweeteningagent such as sucrose or saccharin; or a flavoring agent such aspeppermint, methyl salicylate, or orange flavoring. When the dosage unitform is a capsule, it can contain, in addition to material of the abovetype, a liquid carrier such as a fatty oil. In addition, dosage unitforms can contain various other materials which modify the physical formof the dosage unit, for example, coatings of sugar, shellac, or entericagents.

The active compound or pharmaceutically acceptable salt or derivativethereof can be administered as a component of an elixir, suspension,syrup, wafer, chewing gum or the like. A syrup may contain, in additionto the active compounds, sucrose as a sweetening agent and certainpreservatives, dyes and colorings and flavors.

The active compound or pharmaceutically acceptable derivatives or saltsthereof can also be mixed with other active materials that do not impairthe desired action, or with materials that supplement the desiredaction, such as antibiotics, antifungals, other antiinflammatories, orantiviral compounds.

Solutions or suspensions used for parenteral, intradermal, subcutaneous,or topical application can include the following components: a sterilediluent such as water for injection, saline solution, fixed oils,polyethylene glycols, glycerine, propylene glycol or other syntheticsolvents; antibacterial agents such as benzyl alcohol or methylparabens; antioxidants such as ascorbic acid or sodium bisulfite;chelating agents such as ethylenediaminetetraacetic acid; buffers suchas acetates, citrates or phosphates and agents for the adjustment oftonicity such as sodium chloride or dextrose. The parental preparationcan be enclosed in ampoules, disposable syringes or multiple dose vialsmade of glass or plastic.

If administered intravenously, preferred carriers are physiologicalsaline or phosphate buffered saline (PBS).

In one embodiment, the active compounds are prepared with carriers thatwill protect the compound against rapid elimination from the body, suchas a controlled release formulation, including implants andmicroencapsulated delivery systems.

Biodegradable, biocompatible polymers can be used, such as ethylenevinyl acetate, polyanhydrides, polyglycolic acid, collagen,polyorthoesters, and polylactic acid. Methods for preparation of suchformulations will be apparent to those skilled in the art. The materialscan also be obtained commercially from Alza Corporation (Calif.) andScios Nova (Baltimore, Md.).

Liposomal suspensions may also be pharmaceutically acceptable carriers.These may be prepared accord ng to methods known to those skilled in theart, for example, as described in U.S. Pat. No. 4,522,811 (which isincorporated herein by reference in its entirety). For example, liposomeformulations may be prepared by dissolving appropriate lipid(s) (such asstearoyl phosphatidyl ethanolamine, stearoyl phosphatidyl choline,arachadoyl phosphatidyl choline, and cholesterol) in an inorganicsolvent that is then evaporated, leaving behind a thin film of driedlipid on the surface of the container. An aqueous solution of the activecompound or its monophosphate, diphosphate, and/or triphosphatederivatives are then introduced into the container. The container isthen swirled by hand to free lipid material from the sides of thecontainer and to disperse lipid aggregates, thereby forming theliposomal suspension.

III. Biological Activity

A wide variety of biological assays have also been used to evaluate theability of a compound to inhibit the enzyme 5-lipoxygenase. For example,a cytosol 5-lipoxygenase of rat basophilic leukemia cells (RBL) has beenwidely utilized in studies on leukotriene biosynthesis. Compounds thatinhibit 5-lipoxygenase decrease the levels of leukotrienes.

Another biological assay used to evaluate the ability of a compound toinhibit the enzyme 5-lipoxygenase is based on the classicpharmacological model of inflammation induced by inhibition of LTB, fromionophore stimulated human whole blood.

Any of these assays, or other assays known to those skilled in the art,can be used to evaluate the activity of the compounds disclosed herein.Examples of these evaluations are provided below.

EXAMPLE 2 Effect of Compounds an Cytosol 5-Lipoxygenase of Rat BasophileLeukemia Cells

RBL-2H3 cells were grown to confluence in tissue culture flasksaccording to Carter et al. (J Pharm Exp Ther 256(3); 929-937, 1991). Thecells were harvested and washed five times in calcium-and magnesium-freeD-PBS. The cells were suspended at 2×10⁷/M in 10 mM BES, 10 mM PIPES, pH6.8, 1 mM EDTA and then sonicated. The sonicate was centrifuged at20,000×g for 20 minutes at 4° C. The supernatant was then removed andstored in aliquots at −70° C.

The 5-LO activity in the RBL-2H3 preparation was determined as follows:0.1 ml reactions consisting of 0.7 mM CaCl₂, 100 mM NaCl, 1 mM EDTA, 10mM BES, 10 MM PIPES, pH 7.4, varying concentrations of test compounddissolved in DMSO (7.5% DMSO final in assay), and an amount of theRBL-2H3 preparation that will convert 15% of the arachidonic acidsubstrate mixture to oxygenated products (determined experimentally foreach RBL-2H3preparation), were incubated for 20 minutes at roomtemperature. The reaction was initiated by the addition of 5 μl of thearachidonic acid substrate mixture (0.944 nmol [¹⁴C] arachidonic acidand 6.06 nmol arachidonic acid per assay in 0.028% NH₄OH), and allowedto proceed for 5 minutes at 37° C. The reaction was terminated by theaddition of 0.12 ml of a mixture of (i) 1.66 mg/ml triphenylphosphine inethyl ether; (ii) methanol; and (iii) 0.2M citric acid (30:4:1);followed by centrifugation at 1000×g for 1 minute. 50 μl of the organicphase was drawn into a glass capillary piper and spotted onto silica gel60A TLC plates (Whatman #6KDF). The plates were developed in ethyl etheracetic acid (100:0.1) for 25 minutes at room temperature. The plateswere exposed to Kodak X-OMAT AR film for 40 hours. The film wasdeveloped, scanned using a densitometer, and the peak areas ofarachidonic acid and its product(s) are calculated. The percentinhibition was determined from the amount of [14C]-arachidonic acidconverted into oxygenated products in samples containing test compoundrelative to that of control samples (no test compound).

The results are provided in Table 1.

EXAMPLE 3 Inhibition of Leukotriene B₄, Production inInonophore-stimulated Human Whole Blood

Human blood was drawn into heparinized blood collection tubes, andaliquoted in 1 ml portions into 1.5 ml microfuge tubes. Test compound (5ml) of varying concentrations, dissolved in DMSO, was added to the bloodsample and incubated for 15 minutes at 37° C. Calcium ionophore (5 ml,A23187) in DMSO was added to a final concentration of 50 mM, and thesamples were incubated for 30 minutes at 37° C. Samples are thencentrifuged at 1100×g (2500 rpm, H1000B rotor, in a Sorvall centrifuge)for 10 minutes at 4° C. Supernatant (100 ml) was transferred into a 1.5ml microfuge tube, 400 ml of cold methanol added, and proteinsprecipitated on ice for 30 minutes. The samples were centrifuged at110×g for 10 minutes at 4° C., and the supernatant assayed for LTB₄using a commercially available EIA kit (Cayman Chemical) according tomanufacturer's specifications.

The results are provided in Table 2.

TABLE 2 ex vivo LTB4 RBL HWB dose % time IC50 nM IC50 μm mg/k inh.minutes

1230 1400 0.153   3, iv 3, iv 3, iv   2, po 5, po MOUSE 89 22 36 RAT 3434    15  60 180   900 900

1380 1490  560 1600  720 0.094 0.27 0.078   3, iv 3, iv 3, iv   5, po 2,po 2, po MOUSE 94 22 44 RAT 29 93  5    15  60 180   900  60 360

2000 1260 1560  880 0.43 0.52   2, po 2, po RAT 86 63    60 360

1350 1380 0.15 0.27

 950 0.42

 910 0.18

EXAMPLE 4 Ex-vivo Mouse Whole Blood 5-Lipoxygenase Evaluation

CD-1 female mice, weighing 18-25 grams, and CD female rats, weighing150-230 grams, were obtained from Charles River Labs. Test compoundswere dissolved in 0.5% DMSO in 0.9% NaCl, for administration in mice(0.5 mg/ml) and in an alcohol vehicle (2% benzyl alcohol, 1% ethanol,40% PEG 300, 10% propylene glycol, 47% of 5% dextrose and 3.5% pluronicF-68 in DiH₂O) for use in rats (5 mg/ml). Animals were injected withcompounds (5 mg/kg) or corresponding vehicle (0.5% DMSO in saline, 10ml/kg for mice; alcohol vehicle, 1 ml/kg for rats) 15 minutes beforethey were sacrificed by decapitation. Heparinized whole blood (0.3 ml)was added into 1.5 ml Eppendorf centrifuge tube containing 3 ml of 2 mMcalcium ionophore A23187 (the final concentration of A23187 was 20 mM).The sample was incubated for 30 minutes in a water bath at 37° C., andthen centrifuged for 2 minutes. The plasma was diluted (×120) andassayed for LTB₄ using EIA.

The results are provided in Table 2.

EXAMPLE 5 Glucuronidation Studies

The rate of glucuronidation is a measure of the metabolic stability invivo of the compounds disclosed herein. In vitro glucuronidationreactions were carried out with reaction mixtures containing 2 mg/ml ofhuman microsomal protein, 5 mM magnesium chloride, 100 mM Tris HCl(pH=7.4), 0.1-1.0 mM substrate and 3 mM UDP-glucuronic acid. Afterincubation at 37° C. for 0 (control), 15, 30, 45, 60, 90, 120, 130, and140 minutes 40 μl aliquots of the reaction mixture were mixed with 80 μlof acetonitrile and centrifuged to remove the precipitated protein.Aliquots of the supernatant were analyzed by reverse phase HPLC todetermine the disappearance of parent compounds and formation ofmetabolites. The results are provided in FIG. 2.

EXAMPLE 6 Eosinophil Infiltration Assay

Accumulation of inflammatory cells in the lung is one of thepathological features of asthma. Elevation of leukocytes, particularlyof eosinophils, in blood and lung lavage fluid has been observed afterallergen inhalation in the patients. Eosinophils appear to be importanteffector cells in allergic inflammation, with the cytotoxic propertiesof its granule proteins and the potential of releasing inflammatorymediators. Prevention of allergen-induced eosinophil influx into thelung is considered a credible target for novel anti-asthmatic drugs.

Leukotrienes are products of the arachidonic acid 5-lipoxygenase (5-LO)pathway. Lipoxygenase metabolites (LTB4,5-oxo-15-hydroxy-eicosatetraenoic acid) have been identified thatpossess potent activity to recruit eosinophils. A 5-LO inhibitor whichis able to block immediate bronchoconstriction and also to reduce lateraccumulation of eosinophils into lung tissue consequent to allergenchallenge may be beneficial to the prevention and treatment of asthma.

Eosinophil infiltration into the lung can be measured by counting thecell number in the bronchoalveolar lavage fluid (BALF) fromallergen-challenged guinea-pigs or mice.

Guinea pig model: Female Hartley guinea-pigs, weighing 400-500 g, wereactively sensitized to ovalbumin (OVA) by i.p. injection of 20 μg OVAand 100 mg Al(OH)₃ in 0.5 ml 0.9% NaCl on Day 1 and Day 2. Animals werechallenged with 0.5% OVA (in 0.9% NaCl) aerosol for 30 sec on Day 15 andDay 16. The compounds were prepared in 10% PEG 200 or 0.5%carboxymethylcellulose and administered p.o. 3 times (1 hr. before eachchallenge and between the two challenges). To prevent histaminerelease-induced death, pyrilamine (3 mg/kg, i.p.) was given 15 minutesbefore each challenge. After 24 hours following the first challenge (or4 hours after the last challenge), animals were bled from the carotidunder anesthesia. &AL was performed with 2×10 ml of 0.5 mM EDTA in DPBS(w/o Ca²⁺, Mg²⁺) at 37° C. via a trachea cannulation. The total cells inBAL fluid were measured by a Sysmex microcellcounter (F-800) and thedifferential cells were counted on a cytospin preparation. Percent ofInhibition on total cell or eosinophilaccumulation=[(vehicle—sham)−(treated—sham)]/(vehicle-sham)×100

Mice model: Male C57 BL/6 mice, weighing 21-23 g, were activelysensitized to OVA by administering 10 μg OVA and 1 mg Al(OH)₃ in 0.2 ml0.9% NaCl on Day 1. Hypersensitivity was developed following a dailyinhalation of aerosolized 1% CVA or saline for 30 minutes on Day 14 toDay 21. The compounds were prepared in a 10% PEG 200 or 0.5%carboxymethylcellulose and administered at 20 mg/kg orally, b.i.d. onDay 18 to Day 22. Animals were bled from the carotid under anesthesiafour hours after the last inhalation of OVA. BAL was performed with 2×1ml DPBS (w/o 4 C₈ ²⁺, M₈ ²⁺) containing 0.5 mM sodium EDTA at 37° C. viaa tracheal cannulation. The total cells in BAL fluid were counted by aSysmex microcelcounter (F-800). The differential cells in BAL fluid werecounted by a Sysmex microcellcounter (F-800). The differential cellswere counted on a cytospin preparation and white giemsa stain. % ofInhibition on total cell or Eosinophilaccumulation=[(vehicle—sham)−(treated—sham)]/(vehicle-sham)×100. (SeeYeadon M. et al. Agents Actions 38:8-17, 1993; Brusselle G. G. et al.ALA'94, A754; Schwenk U. et al. J. Biol. Biochem. 267:12482-12488, 1992;and Clinic M. et al. Cur. Opin. Immunol. 6:860-7864, 1994).

Modifications and variations of the present invention relating tocompounds that reduce the formation of oxygen radicals during aninflammatory or immune response will be obvious to those skilled in theart from the foregoing detailed description of the invention. Suchmodifications and variations are intended to come within the scope ofthe appended claims.

We claim:
 1. A method for the treatment of inflammatory bowel disease,comprising administering to a host suffering from or susceptible toinflammatory bowel disease a compound of the following formula:

wherein Ar is aryl or heteroaryl group this is optionally substitutedwith halo, lower alkoxy, lower aryloxy, W, cyano, or R³; m is 0 or 1; nis 1-6; W is independently —AN(OM)C(O)N(R³)R⁴, —N(OM)C(O)N(R³)R⁴,—AN(R³)C(O)N(OM)R⁴, —C(O)N(OM)R⁴, —C(O)NHA; A is lower alkyl, loweralkenyl, lower alkynyl, lower alkylene, lower alkenylene, loweralkynylene, alkylaryl, arylalkyl, alkylarylene, or arylalkylene whereinone or more carbons optionally can be replaced by O, N or S (withvalence completed with hydrogen or oxygen as necessary), provided that—Y—A—, —A—, or —AW— do not include two adjacent heteroatoms; M ishydrogen, a pharmaceutically acceptable cation, or a metabolicallycleavable leaving group; X is O, S, S(O), S(O)₂, NR³, or CHR⁵; Y is O,S, S(O), S(O)₂, NR³, or CHR⁵; Z is O, S, S(O), S(O)₂, or NR³; R¹ and R²are independently hydrogen, lower alkyl, C₃₋₈ cycloalkyl, halo loweralkyl, halo or —COOH; R³ and R⁴ are independently hydrogen, alkyl,alkenyl, alkynyl, aryl, arylalkyl, alkylaryl, C₁₋₆alkoxy-C₁₋₁₀alkyl,C₁₋₆alkylthio-C₁₋₁₀alkyl, heteroaryl, or heteroarylalkyl; and R⁵ ishydrogen, lower alkyl, lower alkenyl, lower alkynyl, arylalkyl,alkylaryl, —AN(OM)C(O)N(R³)R⁴, —AN(R³)C(O)N(OM)R⁴, —AN(OM)C(O)R⁴,—AC(O)N(OM)R⁴, —AS(O)_(x)R³, —AS(O)_(x)CH₂C(O)R³, —AS(O)_(x)CH₂CH(OH)R³,or —AC(O)NHR³, wherein x is 0-2.
 2. The method of claim 1 wherein Ar isphenyl, trimethoxyphenyl, dimethoxyphenyl, fluorophenyl, difluorophenyl,pyridyl, dimethoxypyridyl, quinolinyl, furyl, imidazolyl, or thienyl. 3.The method of claim 1 wherein Ar is 4-fluorophenyl.
 4. The method ofclaim 1 wherein Z is oxygen.
 5. The method of claim 1 wherein Z issulfur.
 6. The method of claim 1 wherein —(Y)_(m)W is selected from thegroup consisting of:


7. The method of claim 1 wherein —(Y)_(m)W is selected from the groupconsisting of:


8. The method of claim 1 wherein the compound is administered in atleast 97% enantiomerically enriched form.
 9. The method of claim 1wherein the compound is selected from the group consisting of2S,5S-trans-2-(4-fluorophenoxymethyl)-5-(4-protected-oxy-1-butynyl)tetrahydrofuranand2S,5R-cis-2-(4-fluorophenoxymethyl)-5-(4-protected-oxy-1-butynyl)tetrahydrofuran.10. The method of claim 1 wherein the compound is2S,5R-cis-2-(4-fluorophenoxymethyl)-5-(4-N-hydroxyureidyl-1-butynyl)tetrahydrofuran.11. The method of claim 1 wherein the compound is2R,5S-cis-2-(4-fluorophenoxymethyl)-5-(4-N-hydroxyureidyl-1-butynyl)tetrahydrofuran.12. The method of claim 1 wherein the compound is2R,5R-trans-2-(4-fluorophenoxymethyl)-5-(4-N-hydroxyureidyl-1-butynyl)tetrahydrofuran.13. The method of claim 1 wherein the compound is2S,5S-trans-2-(4-fluorophenoxymethyl)-5-(4-N-hydroxyureidyl-1-butynyl)tetrahydrofuran.14. The method of claim 1 wherein the compound is selected from thegroup consisting of:trans-2-(3,4,5-trimethoxyphenoxymethyl)-5-(4-N′-methyl-N′-hydroxyureidyl-1-butyl)tetrahydrofuran;trans-2-(3,4,5-trimethoxyphenoxymethyl)-5-(4-N′-methyl-N′-hydroxyureidyl-1-butynyl)tetrahydrofuran;trans-2-(4-fluorophenoxymethyl)-5-(4-N′-methyl-N′-hydroxyureidyl-1-butyl)tetrahydrofuran;trans-2-(4-fluorophenoxymethyl)-5-(4-N′-methyl-N′-hydroxyureidyl-1-butynyl)tetrahydrofuran;trans-2-(4-fluorophenoxymethyl)-5-(4-N′-butyl-N′-hydroxyureidyl-1-butyl)tetrahydrofuran;trans-2-(4-fluorophenoxymethyl)-5-(4-N′-butyl-N′-hydroxyureidyl-1-butynyl)tetrahydrofuran;trans-2-(3,4,5-trimethoxyphenoxymethyl)-5-(4-N′-methyl-N′-hydroxyureidyl-1-butyl)tetrahydrothiophene;trans-2-(3,4,5-trimethoxyphenoxymethyl)-5-(4-N′-methyl-N′-hydroxyureidyl-1-butynyl)tetrahydrothiophene;trans-2-(4-fluorophenoxymethyl)-5-(4-N′-methyl-N′-hydroxyureidyl-1-butyl)tetrahydrothiophene;trans-2-(4-fluorophenoxymethyl)-5-(4-N′-methyl-N′-hydroxyureidyl-1-butynyl)tetrahydrothiophene;trans-2-(4-fluorophenoxymethyl)-5-(4-N′-butyl-N′-hydroxyureidyl-1-butyl)tetrahydrothiophene;andtrans-2-(4-fluorophenoxymethyl)-5-(4-N′-butyl-N′-hydroxyureidyl-1-butynyl)tetrahydrothiophene;and pharmaceutically acceptable salts thereof.
 15. The method of claim 1wherein the host is suffering from or susceptible to Crohn's disease.16. The method of claim 13 wherein the host is suffering from orsusceptible to Crohn's disease.
 17. The method of claim 1 wherein thehost is a mammal, an equine, a canine or a bovine.
 18. The method ofclaim 1 wherein the host is a human.
 19. The method of claim 13 whereinthe host is a mammal, an equine, a canine or a bovine.
 20. The method ofclaim 13 wherein the host is a mammal, an equine, a canine, a bovine ora human.
 21. A method for the treatment of inflammatory bowel disease,comprising administering to a host suffering from or susceptible toinflammatory bowel disease a compound of the following formula:

wherein Ar is aryl or heteroaryl group this is optionally substitutedwith halo, lower alkoxy, lower aryloxy, W, cyano, or R³; m is 0 or 1; nis 1-6; W is independently —N(R³)C(O)N(OM)R⁴, —AN(OM)C(O)R⁴,—N(OM)C(O)R⁴, —AC(O)N(OM)R⁴; A is lower alkyl, lower alkenyl, loweralkynyl, lower alkylene, lower alkenylene, lower alkynylene, alkylaryl,arylalkyl, alkylarylene, or arylalkylene wherein one or more carbonsoptionally can be replaced by O, N or S (with valence completed withhydrogen or oxygen as necessary), provided that —Y—A—, —A—, or —AW— donot include two adjacent heteroatoms, M is hydrogen, a pharmaceuticallyacceptable cation, or a metabolically cleavable leaving group; X is O,S, S(O), S(O)₂, NR³, or CHR⁵; Y is O, S, S(O), S(O)₂, NR³, or CHR⁵; Z isO, S, S(O), S(O)₂, or NR³; R¹ and R² are independently hydrogen, loweralkyl, C₃₋₈ cycloalkyl, halo lower alkyl, halo or —COOH; R³ and R⁴ areindependently hydrogen, alkyl, alkenyl, alkynyl, aryl, arylalkyl,alkylaryl, C₁₋₆alkoxy-C₁₋₁₀alkyl, C₁₋₆alkylthio-C₁₋₁₀alkyl, heteroaryl,or heteroarylalkyl; and R⁵ is hydrogen, lower alkyl, lower alkenyl,lower alkynyl, arylalkyl, alkylaryl, —AN(OM)C(O)N(R³)R⁴,—AN(R³)C(O)N(OM)R⁴, —AN(OM)C(O)R⁴, —AC(O)N(OM)R⁴, —AS(O)_(x)R³,—AS(O)_(x)CH₂C(O)R³, —AS(O)_(x)CH₂CH(OH)R³, or —AC(O)NHR³, wherein x is0-2.